| Toxoplasma gondii is an intracellular parasitic protozoan that can infect nucleated cells of almost all warm-blooded animals.It has a wide range of hosts,with cats being its final host,and intermediate hosts including mammals,birds,reptiles,fish and more than 200 species of animals.Toxoplasmosis is a zoonotic parasitic disease caused by T.gondii infecting the host.It is prevalent worldwide.According to statistics,about 30%of people worldwide are infected with Toxoplasma.In humans,healthy people with Toxoplasma gondii infection may not have obvious symptoms,but severe symptoms may occur when they infect pregnant women,people with weakened immune systems and deficiencies.In animals,T.gondii can cause tissue infections,serious diseases,and even death,which seriously affects livestock production and food safety.The prevention of Toxoplasma gondii is based on early detection,and there are many detection methods.However,due to the diverse genotypes of Toxoplasma gondii and numerous surface antigens,it is very easy to cause missed detection and false positives,which hinders the prevention and control of toxoplasmosis.Therefore,rapid and accurate detection methods for toxoplasmosis have important research significance.Modified agglutination test(MAT)is an internationally recognized gold standard for Toxoplasma detection.It has many advantages such as high accuracy,good specificity,and simple method.It is one of the most commonly used detection methods.However,due to the monopoly of foreign markets,the cost of imported MAT kits is high and the timeliness is poor,and there are still no commercial MAT kits for sale in China.In this experiment,the MAT kit was first prepared and improved,and the kit's antigen type,insect body density,antibody titer,negative and positive criteria,reaction optimal p H and time were explored and improved,and the kit The detection effect,specificity,stability and other aspects of the test were tested.It is determined that the strain of the kit as the antigen is the RH strain.The worm density is 2.5×10~8,the antibody titer is 1:2~8,the optimal p H is 8.5?9,the optimal reaction time is 15 hours to 24 hours,and the negative and positive judgment criteria are performed.Specific experiments were carried out using other parasite infection sera.91 human sera,290 pig sera,and 63 mice were used to test their effects and compared with ELISA and imported MAT kits.At the same time,the repeatability and The stability was evaluated,and the results showed that the prepared MAT kit has a good response to the Toxoplasma infection serum,no crossover,the detection rate is higher than that of ELISA,the accuracy rate is the same as the imported MAT,it can be repeated,and can be stored for a long time.The prepared MAT kit was used to conduct an epidemiological survey of Toxoplasma gondii on 998 human sera from Changchun and 512 human sera from Daqing.The results showed that the infection rate of T.gondii was 23.8%in Changchun and 24.2%in Daqing.Both are higher than the infection rate of T.gondii in the earlier national general survey of parasitic diseases.The actual infection status of T.gondii in Chinese may have increased significantly,which requires attention.This experiment successfully prepared a MAT kit with high accuracy,good specificity,low cost and simple operation,which is of great significance for the rapid clinical detection and prevention of toxoplasmosis. |
PDF Full Text Request